First Sign® Drug of Abuse Buprenorphine Cup Test, First Sign® Drug of Abuse Buprenorphine Dip Card Test, First Sign® Drug of Abuse Butalbital Cup Test, First Sign® Drug of Abuse Butalbital Dip Card Test, First Sign® Drug of Abuse Morphine Cup Test, First Sign® Drug of Abuse Morphine Dip Card Test.
Device Facts
| Record ID | K152551 |
|---|---|
| Device Name | First Sign® Drug of Abuse Buprenorphine Cup Test, First Sign® Drug of Abuse Buprenorphine Dip Card Test, First Sign® Drug of Abuse Butalbital Cup Test, First Sign® Drug of Abuse Butalbital Dip Card Test, First Sign® Drug of Abuse Morphine Cup Test, First Sign® Drug of Abuse Morphine Dip Card Test. |
| Applicant | W.H.P.M., Inc. |
| Product Code | DJG · Clinical Toxicology |
| Decision Date | Nov 9, 2015 |
| Decision | SESE |
| Submission Type | Traditional |
| Regulation | 21 CFR 862.3650 |
| Device Class | Class 2 |
Intended Use
First Sign® Drug of Abuse Buprenorphine Cup Test First Sign® Drug of Abuse Buprenorphine Dip Card Test First Sign® Drug of Abuse Buprenorphine Tests are immunochromatographic assays for the qualitative determination of Buprenorphine, in human urine at cut-off concentration of 10 ng/mL. The tests are available in a Cup format and a Din Card format. The tests may yield preliminary positive results even when prescription drug Buprenorphine is ingescribed doses; it is not intended to distinguish between prescription use or abuse of this drug. There is no uniformly recognized cutoff concentration level for Buprenorphine in urine. The tests provide only preliminary test results. A more specific alternative chemical method must be used in order to obtain a confirmed analytical result. Gas Chromatography/Mass Spectrometry is the preferred confirmatory method. Clinical consideration and professional judgment should be exercised with any drug of abuse test result, particularly when the preliminary result is positive. For in vitro diagnostic use only. The tests are intended for over-the-counter and for prescription use. First Sign® Drug of Abuse Butalbital Cup Test First Sign® Drug of Abuse Butalbital Dip Card Test First Sign® Drug of Abuse Butalbital Tests are immunochromatographic assays for the qualitative determination of Butalbital in human urine at cut-off concentration of 300 ng/mL. The tests are available in a Cup format and a Dip Card format. The tests may yield preliminary positive results even when prescription drug Butalbital is ingested, at prescribed doses; it is not intended to distinguish between prescription use or abuse of this drug. There is no uniformly recognized cutoff concentration level for Butalbital in urine. The tests provide only preliminary test results. A more specific alternative chemical method must be used in order to obtain a confirmed analytical result. Gas Chromatography/Mass Spectrometry is the preferred confirmatory method. Clinical consideration and professional judgment should be exercised with any drug of abuse test result, particularly when the preliminary result is positive. For in vitro diagnostic use only. The tests are intended for over-the-counter and for prescription use. First Sign® Drug of Abuse Morphine Cup Test First Sign® Drug of Abuse Morphine Dip Card Test First Sign® Drug of Abuse Morphine Tests are immunochromatographic assays for the qualitative determination of Morphine in human urine at cut-off concentration of 300 ng/mL. The tests are available in a Cup format and a Dip Card format. The tests provide only preliminary test results. A more specific alternative chemical method must be used in order to obtain a confirmed analytical result. Gas Chromatography/Mass Spectrometry is the preferred confirmatory method. Clinical consideration and professional judgment should be exercised with any drug of abuse test result, particularly when the preliminary result is positive. For in vitro diagnostic use only. The tests are intended for over-the-counter and for prescription use.
Device Story
Lateral flow immunochromatographic assays for qualitative detection of Buprenorphine, Butalbital, or Morphine in human urine; available in cup or dip card formats. Principle: competitive binding; target drug in urine competes with drug-conjugate for limited antibody binding sites on particles. Absence of colored line at test region indicates presence of drug above cut-off; presence of line indicates drug below cut-off. Control line confirms proper flow. Used in point-of-care or home settings by lay users or professionals. Provides preliminary results; requires confirmation by GC/MS. Results assist clinical decision-making regarding drug use status.
Clinical Evidence
Bench testing only. Precision, cut-off verification, interference, and specificity studies performed. Method comparison against GC/MS conducted with 80 clinical samples per test. Lay-user study with 280 participants per test demonstrated high accuracy (95-100%) across various drug concentrations relative to GC/MS. No clinical studies required.
Technological Characteristics
Lateral flow chromatographic immunoassay; competitive binding principle. Monoclonal mouse antibodies. Formats: Cup or Dip Card. Stability: 24 months at 39-86°F. No electronic components or software algorithms.
Indications for Use
Indicated for qualitative detection of Buprenorphine (10 ng/mL), Butalbital (300 ng/mL), or Morphine (300 ng/mL) in human urine. Intended for over-the-counter and prescription use. Not intended to distinguish between prescription use and abuse.
Regulatory Classification
Identification
An opiate test system is a device intended to measure any of the addictive narcotic pain-relieving opiate drugs in blood, serum, urine, gastric contents, and saliva. An opiate is any natural or synthetic drug that has morphine-like pharmocological actions. The opiates include drugs such as morphine, morphine glucoronide, heroin, codeine, nalorphine, and meperedine. Measurements obtained by this device are used in the diagnosis and treatment of opiate use or overdose and in monitoring the levels of opiate administration to ensure appropriate therapy.
Special Controls
*Classification.* Class II (special controls). An opiate test system is not exempt if it is intended for any use other than employment or insurance testing or is intended for Federal drug testing programs. The device is exempt from the premarket notification procedures in subpart E of part 807 of this chapter subject to the limitations in § 862.9, provided the test system is intended for employment and insurance testing and includes a statement in the labeling that the device is intended solely for use in employment and insurance testing, and does not include devices intended for Federal drug testing programs (*e.g.,* programs run by the Substance Abuse and Mental Health Services Administration (SAMHSA), the Department of Transportation (DOT), and the U.S. military).
Related Devices
- K152025 — AssureTech Buprenorphine Tests (Strip, Panel Dip, Quick Cup, Turn-Key Split Cup), AssureTech Methadone Tests (Strip, Panel Dip, Quick Cup, Turn-Key Split Cup) · Assure Tech. Co., Ltd. · Oct 5, 2015
- K150791 — Healgen Secobarbital Test(Strip, Cassette, Cup, Dip Card), Healgen Burprenorphine Test (Strip, Cassette, Cup, Dip Card), Healgen Methadone Test (Strip, Cassette, Cup, Dip Card) · Healgen Scientific,, LLC · Apr 24, 2015
Submission Summary (Full Text)
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Image /page/0/Picture/1 description: The image shows the logo for the Department of Health & Human Services - USA. The logo is circular, with the text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" arranged around the perimeter of the circle. In the center of the circle is a stylized image of a caduceus, which is a symbol of medicine.
Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002
November 9, 2015
W.H.P.M., INC. C/O JOE SHIA REGULATORY CONSULTANT 504E DIAMOND AVE., SUITE I GAITHERSBURG MD 20877
Re: K152551 Trade/Device Name: First Sign® Drug of Abuse Buprenorphine Cup Test First Sign® Drug of Abuse Buprenorphine Dip Card Test First Sign® Drug of Abuse Butalbital Cup Test First Sign® Drug of Abuse Butalbital Dip Card Test First Sign® Drug of Abuse Morphine Cup Test First Sign® Drug of Abuse Morphine Dip Card Test Regulation Number: 21 CFR 862.3650 Regulation Name: Opiate test system Regulatory Class: II Product Code: DJG, DIS Dated: September 1, 2015 Received: September 8, 2015
Dear Mr. Joe Shia:
We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.
If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.
Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Parts 801 and 809); medical device reporting (reporting of
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medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.
If you desire specific advice for your device on our labeling regulations (21 CFR Parts 801 and 809). please contact the Division of Industry and Consumer Education at its toll-free number (800) 638 2041 or (301) 796-7100 or at its Internet address
http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to
http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm for the CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance.
You may obtain other general information on your responsibilities under the Act from the Division of Industry and Consumer Education at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address
http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm.
Sincerely yours.
Courtney H. Lias -S
Courtney H. Lias, Ph.D. Director Division of Chemistry and Toxicology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health
Enclosure
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## Indications for Use
510(k) Number (if known) K152551
#### Device Name
First Sign® Drug of Abuse Buprenorphine Cup Test; First Sign® Drug of Abuse Buprenorphine Dip Card Test First Sign® Drug of Abuse Butalbital Cup Test; First Sign® Drug of Abuse Butalbital Dip Card Test First Sign® Drug of Abuse Morphine Cup Test; First Sign® Drug of Abuse Morphine Dip Card Test
#### Indications for Use (Describe)
First Sign® Drug of Abuse Buprenorphine Cup Test
First Sign® Drug of Abuse Buprenorphine Dip Card Test
First Sign™ Drug of Abuse Buprenorphine Tests are immunochromatographic assays for the qualitative determination of Buprenorphine, in human urine at cut-off concentration of 10 ng/mL. The tests are available in a Cup format and a Din Card format.
The tests may yield preliminary positive results even when prescription drug Buprenorphine is ingescribed doses; it is not intended to distinguish between prescription use or abuse of this drug. There is no uniformly recognized cutoff concentration level for Buprenorphine in urine. The tests provide only preliminary test results. A more specific alternative chemical method must be used in order to obtain a confirmed analytical result. Gas Chromatography/Mass Spectrometry is the preferred confirmatory method. Clinical consideration and professional judgment should be exercised with any drug of abuse test result, particularly when the preliminary result is positive.
For in vitro diagnostic use only. The tests are intended for over-the-counter and for prescription use.
First Sign® Drug of Abuse Butalbital Cup Test
First Sign® Drug of Abuse Butalbital Dip Card Test
First Sign™ Drug of Abuse Butalbital Tests are immunochromatographic assays for the qualitative determination of Butalbital in human urine at cut-off concentration of 300 ng/mL. The tests are available in a Cup format and a Dip Card format.
The tests may yield preliminary positive results even when prescription drug Butalbital is ingested, at prescribed doses; it is not intended to distinguish between prescription use or abuse of this drug. There is no uniformly recognized cutoff concentration level for Butalbital in urine. The tests provide only preliminary test results. A more specific alternative chemical method must be used in order to obtain a confirmed analytical result. Gas Chromatography/Mass Spectrometry is the preferred confirmatory method. Clinical consideration and professional judgment should be exercised with any drug of abuse test result, particularly when the preliminary result is positive.
For in vitro diagnostic use only. The tests are intended for over-the-counter and for prescription use.
First Sign® Drug of Abuse Morphine Cup Test
First Sign® Drug of Abuse Morphine Dip Card Test
First Sign™ Drug of Abuse Morphine Tests are immunochromatographic assays for the qualitative determination of Morphine in human urine at cut-off concentration of 300 ng/mL. The tests are available in a Cup format and a Dip Card format.
The tests provide only preliminary test results. A more specific alternative chemical method must be used in order to obtain a confirmed analytical result. Gas Chromatography/Mass Spectrometry is the preferred confirmatory method. Clinical consideration and professional judgment should be exercised with any drug of abuse test result, particularly when the preliminary result is positive.
For in vitro diagnostic use only. The tests are intended for over-the-counter and for prescription use.
Type of Use (Select one or both, as applicable)
> Prescription Use (Part 21 CFR 801 Subpart D)
Over-The-Counter Use (21 CFR 801 Subpart C)
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#### 510(k) SUMMARY
- 1. Date: November 6, 2015
- 2. Submitter W.H.P.M., Inc. 5358 Irwindale Ave. Irwindale, CA 91706
- 3. Contact person: John Wan W.H.P.M., Inc. 5358 Irwindale Ave. Irwindale, CA 91706 Telephone: 626-443-8480 Fax: 626-443-8065 Email: johnwan@whpm.com
- First Sign® Drug of Abuse Buprenorphine Cup Test 4. Device Name: First Sign® Drug of Abuse Buprenorphine Dip Card Test First Sign® Drug of Abuse Butalbital Cup Test First Sign® Drug of Abuse Butalbital Dip Card Test First Sign® Drug of Abuse Morphine Cup Test First Sign® Drug of Abuse Morphine Dip Card Test
- Buprenorphine Urine Test Common Name: Butalbital Urine Test Morphine Urine Test
| Product Code | Class | CFR # | Panel |
|--------------|----------|------------------------------------------|------------|
| DJG | Class II | 21 CFR, 862.3650 Opiate Test System | Toxicology |
| DIS | Class II | 21 CFR, 862.3150 Barbiturate Test System | Toxicology |
| DJG | Class II | 21 CFR, 862.3650 Opiate Test System | Toxicology |
This 510(k) summary is for three drug tests namely. BUP, Butalbital and MOP tests. The performance studies including the lay user study were performed on each of the three durg tests (BUP, Butalbital and MOP tests) separately.
- 5. Predicate Devices:
K122809, Advin Multi Drug Screen Test K041712 RapidTec 4
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#### 6. Intended Use
First Sign® Drug of Abuse Buprenorphine Cup Test First Sign® Drug of Abuse Buprenorphine Dip Card Test First Sign® Drug of Abuse Buprenorphine Tests are immunochromatographic assays for the qualitative determination of Buprenorphine, in human urine at cut-off concentration of 10 ng/mL. The tests are available in a Cup format and a Dip Card format.
The tests may vield preliminary positive results even when prescription drug Buprenorphine is ingested. at prescribed doses; it is not intended to distinguish between prescription use or abuse of this drug. There is no uniformly recognized cutoff concentration level for Buprenorphine in urine. The tests provide only preliminary test results. A more specific alternative chemical method must be used in order to obtain a confirmed analytical result. Gas Chromatography/Mass Spectrometry is the preferred confirmatory method. Clinical consideration and professional judgment should be exercised with any drug of abuse test result, particularly when the preliminary result is positive.
For in vitro diagnostic use only. The tests are intended for over-the-counter and for prescription use.
First Sign® Drug of Abuse Butalbital Cup Test
First Sign® Drug of Abuse Butalbital Dip Card Test
First Sign® Drug of Abuse Butalbital Tests are immunochromatographic assays for the qualitative determination of Butalbital in human urine at cut-off concentration of 300 ng/mL. The tests are available in a Cup format and a Dip Card format.
The tests may vield preliminary positive results even when prescription drug Butalbital is ingested, at prescribed doses; it is not intended to distinguish between prescription use or abuse of this drug. There is no uniformly recognized cutoff concentration level for Butalbital in urine. The tests provide only preliminary test results. A more specific alternative chemical must be used in order to obtain a confirmed analytical result. Gas Chromatography/Mass Spectrometry is the preferred confirmatory method. Clinical consideration and professional judgment should be exercised with any drug of abuse test result, particularly when the preliminary result is positive.
For in vitro diagnostic use only. The tests are intended for over-the-counter and for prescription use.
First Sign® Drug of Abuse Morphine Cup Test
First Sign® Drug of Abuse Morphine Dip Card Test
First Sign® Drug of Abuse Morphine Tests are immunochromatographic assays for the qualitative determination of Morphine in human urine at cut-off concentration of 300 ng/mL. The tests are available in a Cup format and a Dip Card format.
The tests provide only preliminary test results. A more specific alternative chemical method must be used in order to obtain a confirmed analytical result. Gas Chromatography/Mass Spectrometry is the
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preferred confirmatory method. Clinical consideration and professional judgment should be exercised with any drug of abuse test result, particularly when the preliminary result is positive. For in vitro diagnostic use only. The tests are intended for over-the-counter and for prescription use.
- 7. Device Description
First Sign™ Drug of Abuse Buprenorphine Test, First Sign™ Drug of Abuse Butalbital Test and First Sign™ Drug of Abuse Morphine Test are immunochromatographic assays. Each assay test is a lateral flow system for the qualitative detection of Buprenorphine, or Butalbital or Morphine in human urine. The products are single-use in vitro diagnostic devices, which come in the formats of Dip Cards or Cups. Each test kit contains a Test Device (in one of the two formats), a package insert and a urine cup for sample collection. Each test device is sealed with a desiccant in an aluminum pouch.
## 8. Substantial Equivalence Information
A summary comparison of features of the candidate device and the predicate device is provided in Table 1, Table 2 & Table 3.
| Item | Candidate Device<br>First Sign® Drug of Abuse<br>Buprenorphine Test | Predicate Device<br>(k122809)<br>Advin Multi Drug<br>Screen Test |
|--------------------------|-------------------------------------------------------------------------------------------------------------------------------------|------------------------------------------------------------------|
| Indication(s)<br>for Use | For the qualitative determination of<br>Buprenorphine in human urine. | Same |
| Calibrator | Buprenorphine | Same |
| Methodology | Competitive binding, lateral flow<br>immunochromatographic assays based on<br>the principle of antigen antibody<br>immunochemistry. | Same |
| Specimen Type | Human urine | Same |
| Cut-Off Values | 10 ng/mL | Same |
| Intended<br>Population | For over-the-counter and prescription<br>uses. | For<br>over-the-counter<br>use. |
| Configurations | Cup, Dip Card | Cup, Dip Card,<br>Cassette |
Table 1: Features Comparison of First Sign® Drug of Abuse Buprenorphine Test and the Predicate Device
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Table 2: Features Comparison of First Sign® Drug of Abuse Butalbital Test and the Predicate Device
| | Candidate Device | Predicate Device |
|-----------------------|-------------------------------------------------------------------------------------------------------------------------------------|-------------------------------------------------------------------|
| Item | First Sign® Drug of Abuse Butalbital Test | (k041712) RapidTec 4 |
| Indication(s) for Use | For the qualitative determination of Butalbital in human urine. | For the qualitative determination of Barbiturates in human urine. |
| Calibrator | Butalbital | Same |
| Methodology | Competitive binding, lateral flow<br>immunochromatographic assays based on<br>the principle of antigen antibody<br>immunochemistry. | Same |
| Specimen Type | Human urine | Same |
| Cut-Off Values | 300 ng/mL | Same |
| Intended Population | For over-the-counter and prescription uses. | For prescription use. |
| Configurations | Cup, Dip Card | Strip |
Table 3: Features Comparison of First Sign® Drug of Abuse Morphine Test and the Predicate Device
| Item | Candidate Device<br>First Sign® Drug of Abuse Morphine<br>Test | Predicate Device<br>(k122809)<br>Advin Multi Drug<br>Screen Test |
|--------------------------|-------------------------------------------------------------------------------------------------------------------------------------|------------------------------------------------------------------|
| Indication(s)<br>for Use | For the qualitative determination of<br>Morphine in human urine. | Same |
| Calibrator | Morphine | Same |
| Methodology | Competitive binding, lateral flow<br>immunochromatographic assays based on<br>the principle of antigen antibody<br>immunochemistry. | Same |
| Specimen Type | Human Urine | Same |
| Cut-Off Values | 300 ng/mL | Same |
| Intended<br>Population | For over-the-counter and prescription<br>uses. | For<br>over-the-counter<br>use. |
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| Item | Candidate Device<br>First Sign® Drug of Abuse Morphine<br>Test | Predicate Device<br>(k122809)<br>Advin Multi Drug<br>Screen Test |
|----------------|----------------------------------------------------------------|------------------------------------------------------------------|
| Configurations | Cup, Dip Card | Cup, Dip Card,<br>Cassette |
## 9. Test Principle
Each assay test is a lateral flow chromatographic immunoassay. During testing, a urine specimen migrates upward by capillary action. If target drugs are present in the urine specimen below its cut-off concentration, it will not saturate the binding sites of its specific antibody (monoclonal mouse antibody) coated on the particles. The antibody-coated particles will then be captured by immobilized drug-conjugate and a visible colored line will show up in the test line region. The colored line will not form in the test line region if the target drug level exceeds its cut-off concentration because it will saturate all the binding sites of the antibody coated on the particles. A band should form in the control region of the devices regardless of the presence of drug or metabolite in the sample.
## 10. Performance Characteristics
## First Sign® Drug of Abuse Buprenorphine Test
Analytical Performance
- a. Precision
Precision studies were carried out for samples with concentrations of -100% cut-off, -50% cut-off, -25% cut-off, at the cut-off, +25% cut-off, +75% cut-off, +75% cut-off and +100% cut-off. These samples were prepared by spiking drug in negative urine samples. Each drug concentration was confirmed by GC/MS. All sample aliquots were blind-labeled and randomized by the person who prepared samples and did not take part in the sample testing. For each concentration, tests were performed two runs per day for 25 days by three different operators for each format of devices. Different set of operators tested each format. The results obtained are summarized in the following tables:
| | Result | -100%<br>Cut-off | -75%<br>Cut-off | -50%<br>Cut-off | -25%<br>Cut-off | Cut-off | +25%<br>Cut-off | +50%<br>Cut-off | +75%<br>Cut-off | +100%<br>Cut-off |
|------|--------|------------------|-----------------|-----------------|-----------------|---------|-----------------|-----------------|-----------------|------------------|
| Drug | Lot 1 | 50-/0+ | 50-/0+ | 50-/0+ | 50-/0+ | 3-/47+ | 50+/0- | 50+/0- | 50+/0- | 50+/0- |
| | Lot 2 | 50-/0+ | 50-/0+ | 50-/0+ | 50-/0+ | 3-/47+ | 50+/0- | 50+/0- | 50+/0- | 50+/0- |
| | Lot 3 | 50-/0+ | 50-/0+ | 50-/0+ | 50-/0+ | 3-/47+ | 50+/0- | 50+/0- | 50+/0- | 50+/0- |
#### Buprenorphine Dip Card Format
## Buprenorphine Cup Format
| | Result | -100%<br>Cut-off | -75%<br>Cut-off | -50%<br>Cut-off | -25%<br>Cut-off | +25%<br>Cut-off | +50%<br>Cut-off | +75%<br>Cut-off | +100%<br>Cut-off |
|------|--------|------------------|-----------------|-----------------|-----------------|-----------------|-----------------|-----------------|------------------|
| Drug | Lot 4 | 50-/0+ | 50-/0+ | 50-/0+ | 50-/0+ | 3-/47+ | 50+/0- | 50+/0- | 50+/0- |
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| Drug | Result | -100%<br>Cut-off | -75%<br>Cut-off | -50%<br>Cut-off | -25%<br>Cut-off | Cut-off | +25%<br>Cut-off | +50%<br>Cut-off | +75%<br>Cut-off | +100%<br>Cut-off |
|-------|--------|------------------|-----------------|-----------------|-----------------|---------|-----------------|-----------------|-----------------|------------------|
| Lot 5 | | 50-/0+ | 50-/0+ | 50-/0+ | 50-/0+ | 2-/48+ | 50+/0- | 50+/0- | 50+/0- | 50+/0- |
| Lot 6 | | 50-/0+ | 50-/0+ | 50-/0+ | 50-/0+ | 3-/47+ | 50+/0- | 50+/0- | 50+/0- | 50+/0- |
## b. Linearity
Not applicable.
## c. Stability
The devices are stable at 39-86°F (4-30°C) for 24 months based on the accelerated stability study at 50°C. Control materials are not provided with the device. The labeling provides information on how to obtain control materials.
## d. Cut-off
A total of 150 samples equally distributed at concentrations of -50% cut-off; -25% cut-off; cut-off; +25% cut-off; +50% cut-off were tested using three different lots of each device by three different operators. Results were all positive at and above +25% cut-off and all negative at and below -25% cut-off for Buprenorphine. The following cut-off value for the test devices have been verified.
| Test | Calibrator | Cut-off (ng/mL) |
|-------------------------------------------------|---------------|-----------------|
| First Sign® Drug of Abuse<br>Buprenorphine Test | Buprenorphine | 10 |
## e. Interference
Potential interfering substances found in human urine of physiological or pathological conditions were added to drug-free urine and to urine containing target drugs at 25% below and 25% above cut-off levels. These urine samples were tested using three lots of each device for all formats.
Compounds that showed no interference at a concentration of 100ug/mL are summarized in the following tables. There were no differences observed for different formats.
| 4-Acetamidophenol | β-Estradiol | Oxolinic acid |
|-----------------------|--------------------------------------------------------|--------------------------------------------------|
| Acetophenetidin | Estrone-3-sulfate | Oxycodone |
| N-Acetylprocainamide | Ethyl-p-aminobenzoate | Oxymetazoline |
| Acetylsalicylic acid | 2-ethylidene-1,5-dimethyl-3,3<br>- diphenylpyrrolidine | Papaverine |
| Aminopyrine | Fenoprofen | Penicillin-G |
| Amobarbital | Furosemide | Pentazocine hydrochloride |
| Amoxicillin | Gentisic acid | Pentobarbital |
| Amphetamine | Hemoglobin | Perphenazine |
| Ampicillin | Hydralazine | Phencyclidine |
| L-Ascorbic acid | Hydrochlorothiazide | Phenelzine |
| Apomorphine | Hydrocodone | Phenobarbital |
| Aspartame | Hydrocortisone | Phentermine |
| Atropine | O-Hydroxyhippuric acid | β-Phenylethylamine |
| Benzilic acid | p-Hydroxyamphetamine | Trans-2-phenylcyclopropylam<br>ine hydrochloride |
| Benzoic acid | p-Hydroxy-<br>methamphetamine | L-Phenylephrine |
| Benzoylecgonine | 3-Hydroxytyramine | Phenylpropanolamine |
| Benzphetamine | Ibuprofen | Prednisolone |
| Bilirubin | Iprazid | Prednisone |
| (±) - Brompheniramine | (±) - Isoproterenol | Procaine |
| Butalbital | Isoxsuprine | DL-Propranolol |
| Caffeine | Ketamine | D-Propoxyphene |
| Cannabidiol | Ketoprofen | D-Pseudoephedrine |
| Cannabinol | Labetalol | Quinacrine |
| Chloralhydrate | Loperamide | Quinidine |
| Chloramphenicol | Methylenedioxyethylampheta<br>mine | Quinine |
| Chlorothiazide | Meperidine | Ranitidine |
| (±) Chlorpheniramine | Meprobamate | Salicylic acid |
| Chlorpromazine | Methadone | Secobarbital |
| Chlorquine | Methamphetamine | Serotonin |
| Cholesterol | Methoxyphenamine | Sulfamethazine |
| Clonidine | (±)-3,4-Methylenedioxyamph<br>etamine hydrochloride | Sulindac |
| Cocaethylene | Methylenedioxymethampheta<br>mine | Tetracycline |
| Cocaine hydrochloride | Morphine-3-β-Dglucuronide | Tetrahydrocortisone3-(ß-D-gl<br>ucuronide) |
| Codeine | Morphine | Tetrahydrozoline |
| Cortisone | Morphine sulfate | Thiamine |
| (-) Cotinine | 11-nor-Δ9-THC-9-COOH | Thioridazine |
| Creatinine | Nalidixic acid | DL-Tyrosine |
| Deoxycorticosterone | Naloxone | Tolbutamide |
| Dextromethorphan | Naltrexone | Triamterene |
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| Diflunisal | Niacinamide | Trimethoprim |
|------------------------|-------------------|---------------|
| Digoxin | Nifedipine | Tryptamine |
| Diphenhydramine | Norcodeine | DL-Tryptophan |
| Doxylamine | Norethindrone | Tyramine |
| Ecgonine hydrochloride | D-Norpropoxyphene | Uric acid |
| Ecgonine methyl ester | Nortriptyline | Verapamil |
| Ephedrine | Noscapine | Zomepirac |
| (L) - Epinephrine | Oxalic acid | |
| Erythromycin | Oxazepam | |
## f. Specificity
To test the specificity, drug metabolites and other components that are likely to interfere in urine samples were tested using three lots of each device for all formats. The obtained lowest detectable concentration was used to calculate the cross-reactivity. There were no differences observed for different formats.
| Buprenorphine | Result | % |
|----------------------------------|--------------------------|------------------|
| Cut-off=10 ng/mL | | Cross-Reactivity |
| Buprenorphine | Positive at 10 ng/mL | 100% |
| Buprenorphine -3-D-Glucuronide | Positive at 15 ng/mL | 67% |
| Norbuprenorphine | Positive at 40 ng/mL | 25% |
| Norbuprenorphine-3-D-Glucuronide | Positive at 500 ng/mL | 2% |
| Morphine | Negative at 100000 ng/mL | Not Detected |
| Oxymorphone | Negative at 100000 ng/mL | Not Detected |
| Hydromorphone | Negative at 100000 ng/mL | Not Detected |
## g. Effect of Urine Specific Gravity and Urine pH
To investigate the effect of urine specific gravity and urine pH, urine samples with a range of 1.000 to 1.035 specific gravity or urine samples with a range of pH 4 to 9 were spiked with target drugs at 25% below and 25% above cut-off levels. These samples were tested using three lots of each device for all formats. Results were all positive for samples at and above +25% Cut-Off and all negative for samples at and below -25% Cut-Off. There were no differences observed for different formats.
## h. Comparison Studies
The method comparison studies for the First Sign® Drug of Abuse Buprenorphine Test was performed in-house with three different laboratory assistants for each format of the device. Operators ran 80 (40 negative and 40 positive) unaltered clinical samples. The samples were blind labeled and compared to GC/MS results. The results are presented in the tables below:
{12}------------------------------------------------
| Dip Card<br>format | | Negative | Low<br>Negative<br>by GC/MS<br>(less than<br>-50%) | Near Cutoff<br>Negative by<br>GC/MS<br>(Between<br>-50% and<br>cut-off) | Near<br>Cutoff<br>Positive by<br>GC/MS<br>(Between<br>the cut-off<br>and +50%) | High<br>Positive by<br>GC/MS<br>(greater<br>than +50%) |
|--------------------|----------|----------|----------------------------------------------------|-------------------------------------------------------------------------|--------------------------------------------------------------------------------|--------------------------------------------------------|
| Viewer A | Positive | 0 | 0 | 1 | 13 | 26 |
| | Negative | 10 | 10 | 19 | 1 | 0 |
| Viewer B | Positive | 0 | 0 | 1 | 13 | 26 |
| | Negative | 10 | 10 | 19 | 1 | 0 |
| Viewer C | Positive | 0 | 0 | 0 | 13 | 26 |
| | Negative | 10 | 10 | 20 | 1 | 0 |
# Discordant Results
| Viewer | Sample Number | GC/MS (ng/mL) Result | Dipcard Format Viewer Results |
|----------|---------------|----------------------|-------------------------------|
| Viewer A | 2014122568 | 9.5 | Positive |
| Viewer A | 2014122575 | 10.5 | Negative |
| Viewer B | 2014122505 | 10.8 | Negative |
| Viewer B | 2014122327 | 9.4 | Positive |
| Viewer C | 2014122561 | 10.7 | Negative |
| Cup<br>format | | Negative | Low<br>Negative<br>by GC/MS<br>(less than<br>-50%) | Near Cutoff<br>Negative by<br>GC/MS<br>(Between<br>-50% and<br>cut-off) | Near<br>Cutoff<br>Positive by<br>GC/MS<br>(Between<br>the cut-off<br>and +50%) | High<br>Positive by<br>GC/MS<br>(greater<br>than +50%) |
|---------------|----------|----------|----------------------------------------------------|-------------------------------------------------------------------------|--------------------------------------------------------------------------------|--------------------------------------------------------|
| Viewer A | Positive | 0 | 0 | 0 | 13 | 26 |
| | Negative | 10 | 10 | 20 | 1 | 0 |
| Viewer B | Positive | 0 | 0 | 1 | 13 | 26 |
| | Negative | 10 | 10 | 19 | 1 | 0 |
| Viewer C | Positive | 0 | 0 | 1 | 13 | 26 |
| | Negative | 10 | 10 | 19 | 1 | 0 |
# Discordant Results
| Viewer | Sample Number | GC/MS (ng/mL) Result | Cup Format Viewer Results |
|----------|---------------|----------------------|---------------------------|
| Viewer A | 2014122575 | 10.5 | Negative |
| Viewer B | 2014122764 | 9 | Positive |
{13}------------------------------------------------
| Viewer | Sample Number | GC/MS (ng/mL)<br>Result | Cup Format<br>Viewer Results |
|----------|---------------|-------------------------|------------------------------|
| Viewer B | 2014122561 | 10.7 | Negative |
| Viewer C | 2014122568 | 9.5 | Positive |
| Viewer C | 2014122505 | 10.8 | Negative |
## i. Lay-user study
A lay user study was performed at three intended user sites with 280 lay persons testing the Buprenorphine devices. A total of 142 females and 138 males tested the Buprenorphine samples. They had diverse educational and professional backgrounds and ranged in age from 21 to > 50 years. Urine samples were prepared at the following concentrations; negative, +/-75%, +/-50%, +/-25% of the cutoff by spiking drugs into drug free-pooled urine specimens. The concentrations of the samples were confirmed by GC/MS. Each sample was aliquoted into individual containers and blind-labeled. Each participant was provided with the package insert, 1 blind labeled sample and a device. The results are summarized below.
| % of Cutoff | Number<br>of<br>samples | Buprenorphine<br>Concentration by GC/MS<br>(ng/mL) | Lay person results | | The<br>percentage of<br>correct results<br>(%) |
|--------------|-------------------------|----------------------------------------------------|--------------------|--------------------|------------------------------------------------|
| | | | No. of<br>Positive | No. of<br>Negative | |
| -100% Cutoff | 20 | 0 | 0 | 20 | 100% |
| -75% Cutoff | 20 | 2.6 | 0 | 20 | 100% |
| -50% Cutoff | 20 | 5.2 | 0 | 20 | 100% |
| -25% Cutoff | 20 | 7.8 | 0 | 20 | 100% |
| +25% Cutoff | 20 | 13 | 19 | 1 | 95% |
| +50% Cutoff | 20 | 15.7 | 20 | 0 | 100% |
| +75% Cutoff | 20 | 18.3 | 20 | 0 | 100% |
Comparison between GC/MS and Lay Person Results (Buprenorphine DipCard)
Comparison between GC/MS and Lay Person Results (Buprenorphine Cup)
| | Number | Buprenorphine | Lay person results | | The | |
|-------------|---------------|-----------------------------------|------------------------------------------|----|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|--|
| % of Cutoff | of<br>samples | Concentration by GC/MS<br>(ng/mL) | No. of<br>No. of<br>Positive<br>Negative | | percentage of<br>correct results<br>(%) | |
| -100%Cutoff | 20 | 0 | 0 | 20 | 100% | |
| -75%Cutoff | 20 | 2.6 | 0 | 20 | 100% | |
| -50% Cutoff | 20 | 5.2 | 0 | 20 | 100% | |
| -25% Cutoff | 20 | 7.8 | 1 | 19 | તે તે જે જ જીરી તેમ જ દૂધની ડેરી જેવી સવલતો પ્રાપ્ય થયેલી છે. આ ગામનાં પ્રાથમિક શાળા, પંચાયતઘર, આંગણવાડી તેમ જ દૂધની ડેરી જેવી સવલતો પ્રાપ્ય થયેલી છે. આ ગામનાં લોકોનો મુખ્ય | |
| +25% Cutoff | 20 | 13 | 19 | 1 | તે તે જે જ જીરી તેમ જ દૂધની ડેરી જેવી સવલતો પ્રાપ્ય થયેલી છે. આ ગામનાં પ્રાથમિક શાળા, પંચાયતઘર, આંગણવાડી તેમ જ દૂધની ડેરી જેવી સવલતો પ્રાપ્ય થયેલી છે. આ ગામનાં લોકોનો મુખ્ય | |
| +50% Cutoff | 20 | 15.7 | 20 | 0 | 100% | |
| +75% Cutoff | 20 | 18.3 | 20 | 0 | 100% | |
{14}------------------------------------------------
Lay-users were also given surveys on the ease of understanding the package insert instructions. All lay users indicated that the device instructions can be easily followed. A Flesch-Kincaid reading analysis was performed on each package insert and the scores revealed a reading Grade Level of 7.
- j. Clinical Studies Not applicable.
## First Sign® Drug of Abuse Butalbital Test
Analytical Performance
- a. Precision
Precision studies were carried out for samples with concentrations of -100% cut-off, -50% cut-off, -25% cut-off, at the cut-off, +25% cut-off, +75% cut-off, +75% cut-off and +100% cut-off. These samples were prepared by spiking drug in negative urine samples. Each drug concentration was confirmed by GC/MS. All sample aliquots were blind-labeled and randomized by the person who prepared samples and did not take part in the sample testing. For each concentration, tests were performed two runs per day for 25 days by three different operators for each format of devices. Different set of operators tested each format. The results obtained are summarized in the following tables:
| | Result | -100%<br>Cut-off | -75%<br>Cut-off | -50%<br>Cut-off | -25%<br>Cut-off | Cut-off | +25%<br>Cut-off | +50%<br>Cut-off | +75%<br>Cut-off | +100%<br>Cut-off |
|-------|--------|------------------|-----------------|-----------------|-----------------|---------|-----------------|-----------------|-----------------|------------------|
| Drug | | | | | | | | | | |
| Lot 1 | | 50-/0+ | 50-/0+ | 50-/0+ | 50-/0+ | 3-/47+ | 50+/0- | 50+/0- | 50+/0- | 50+/0- |
| Lot 2 | | 50-/0+ | 50-/0+ | 50-/0+ | 50-/0+ | 4-/46+ | 50+/0- | 50+/0- | 50+/0- | 50+/0- |
| Lot 3 | | 50-/0+ | 50-/0+ | 50-/0+ | 50-/0+ | 3-/47+ | 50+/0- | 50+/0- | 50+/0- | 50+/0- |
#### Butalbital Dip Card Format
## Butalbital Cup Format
| | Result | -100%<br>Cut-off | -75%<br>Cut-off | -50%<br>Cut-off | -25%<br>Cut-off | Cut-off | +25%<br>Cut-off | +50%<br>Cut-off | +75%<br>Cut-off | +100%<br>Cut-off |
|------|--------|------------------|-----------------|-----------------|-----------------|---------|-----------------|-----------------|-----------------|------------------|
| Drug | Lot 4 | 50-/0+ | 50-/0+ | 50-/0+ | 50-/0+ | 3-/47+ | 50+/0- | 50+/0- | 50+/0- | 50+/0- |
| | Lot 5 | 50-/0+ | 50-/0+ | 50-/0+ | 50-/0+ | 3-/47+ | 50+/0- | 50+/0- | 50+/0- | 50+/0- |
| | Lot 6 | 50-/0+ | 50-/0+ | 50-/0+ | 50-/0+ | 3-/47+ | 50+/0- | 50+/0- | 50+/0- | 50+/0- |
#### b. Linearity
Not applicable.
{15}------------------------------------------------
- c. Stability
The devices are stable at 39-86°F (4-30°C) for 24 months based on the accelerated stability study at 50°C. Control materials are not provided with the device. The labeling provides information on how to obtain control materials.
- d. Cut-off
A total of 150 samples equally distributed at concentrations of -50% cut-off; -25% cut-off; cut-off; +25% cut-off; +50% cut-off were tested using three different lots of each device by three different operators. Results were all positive at and above +25% cut-off and all negative at and below -25% cut-off for Butalbital. The following cut-off value for the test devices have been verified.
| Test | Calibrator | Cut-off (ng/mL) |
|----------------------------------------------|------------|-----------------|
| First Sign® Drug of Abuse Butalbital<br>Test | Butalbital | 300 |
## e. Interference
Potential interfering substances found in human urine of physiological conditions were added to drug-free urine and to urine containing target drugs at 25% below and 25% above cut-off levels. These urine samples were tested using three lots of each device for all formats.
Compounds that showed no interference at a concentration of 100µg/mL are summarized in the following tables. There were no differences observed for different formats.
| Acetaminophen | Erythromycin | 11-nor-Δ9-THC-9-COOH |
|-----------------------------------------------------|---------------------------------------------------|------------------------|
| Acetophenetidin | β-Estradiol | O-Hydroxyhippuric acid |
| Acetylsalicylic acid | Estrone-3-sulfate | D,L-Octopamine |
| Aminopyrine | Ethyl-p-aminobenzoate | Oxalic acid |
| Amitriptyline | Fenoprofen | Oxazepam |
| Amoxicillin | Furosemide | Oxolinic acid |
| Amphetamine | Gentisic acid | Oxycodone |
| Ampicillin | Hemoglobin | Oxymetazoline |
| Apomorphine | Hydralazine | Papaverine |
| Ascorbic acid | Hydrochlorothiazide | Penicillin-G |
| Aspartame | Hydrocodone | Pentazocaine |
| Atropine | Hydrocortisone | Perphenazine |
| Benzilic acid | p-Hydroxyamphetamine | Phencyclidine |
| Benzoic acid | p-Hydroxymethamphetamine | Phenelzine |
| Benzoylecgonine | 3-Hydroxytyramine | β-Phenylethlamine |
| Bilirubin | Ibuprofen | Phenylpropanolamine |
| Brompheniramine | Imipramine | Prednisolone |
| Buprenorphine | (-) Isoproterenol | Prednisone |
| Caffeine | Isoxsuprine | Procaine |
| Cannabidiol | Ketamine | Promazine |
| Cannabinol | Ketoprofen | Promethazine |
| Chloralhydrate | Labetalol | D,L-Propanolol |
| Chloramphenicol | Levorphanol | D-Propoxyphene |
| Chlorothiazide | Loperamide | Quinidine |
| (±)Chlorpheniramine | L-Phenylephrine | Quinine |
| Chlorpromazine | Maprotiline | Ranitidine |
| Chlorquine | Meperidine | Salicylic acid |
| Cholesterol | Meprobamate | Serotonin |
| Clomipramine | Morphine-3-β-D glucuronide | Sulfamethazine |
| Clonidine | Methadone | Sulindac |
| Cocaine hydrochloride | Methamphetamine | Temazepam |
| Codeine | (±)-3,4-Methylenedioxy- amphetamine hydrochloride | Tetracycline |
| Cortisone | Methylenedioxy- methamphetamine | Tetrahydrozoline |
| (-) Cotinine | Morphine | Thebaine |
| Creatinine | Morphine Sulfate | Thiamine |
| Deoxycorticosterone | N-Acetylprocainamide | Thioridazine |
| Dextromethorphan | Nalidixic acid | Triamterene |
| Diazepam | Naloxone | Trifluoperazine |
| Diclofenac | Naltrexone | Trimethoprim |
| Diflunisal | Naproxen | Trimipramine |
| Digoxin | Niacinamide | Tryptamine |
| Diphenhydramine | Nifedipine | D, L-Tyrosine |
| Doxylamine | Norcodein | Uric acid |
| Ecgonine hydrochloride | Norethindrone | Verapamil |
| Ecgonine methylester | D-Norpropoxyphene | Zomepirac |
| (IR,2S)(-)Ephedrine | Noscapine | |
| 2-ethylidene-1,5-dimethyl-3, 3- diphenylpyrrolidine | Nortriptyline | |
{16}------------------------------------------------
#### f. Specificity
To test the specificity, drug metabolites and other components that are likely to interfere in urine samples were tested using three lots of each device for all formats. The obtained lowest detectable concentration was used to calculate the cross-reactivity. There were no differences observed for different formats.
{17}------------------------------------------------
| Butalbital | Result | % Cross-Reactivity |
|--------------------|------------------------|--------------------|
| Cut-off=300 ng/mL) | | |
| Butalbital | Positive at 300 ng/mL | 100% |
| Secobarbital | Positive at 300 ng/mL | 100% |
| Amobarbital | Positive at 3000 ng/mL | 10% |
| Alphenal | Positive at 250 ng/mL | 120% |
| Aprobarbital | Positive at 200 ng/mL | 150% |
| Allobarbital | Positive at 500 ng/mL | 60% |
| Butabarbital | Positive at 1000 ng/mL | 30% |
| Butethal | Positive at 500 ng/mL | 60% |
| Cyclopentobarbital | Positive at 300 ng/mL | 100% |
| Pentobarbital | Positive at 1300 ng/mL | 23% |
| Phenobarbital | Positive at 1900 ng/mL | 16% |
g. Effect of Urine Specific Gravity and Urine pH
To investigate the effect of urine specific gravity and urine pH, urine samples with a range of 1.000 to 1.035 specific gravity or urine samples with a range of pH 4 to 9 were spiked with target drugs at 25% below and 25% above cut-off levels. These samples were tested using three lots of each device for all formats. Results were all positive for samples at and above +25% Cut-Off and all negative for samples at and below -25% Cut-Off. There were no differences observed for different formats.
- h. Comparison Studies
The method comparison studies for the First Sign® Drug of Abuse Butalbital Test was performed in-house with three different laboratory assistants for each format of the device. Operators ran 80 (40 negative and 40 positive) unaltered clinical samples. The samples were blind labeled and compared to GC/MS results. The results are presented in the tables below:
| Dip Card<br>format | | Negative | Low<br>Negative<br>by GC/MS<br>(less than<br>-50%) | Near Cutoff<br>Negative by<br>GC/MS<br>(Between<br>-50% and<br>cut-off) | Near<br>Cutoff<br>Positive by<br>GC/MS<br>(Between<br>the cut-off<br>and +50%) | High<br>Positive by<br>GC/MS<br>(greater<br>than +50%) |
|--------------------|----------|----------|----------------------------------------------------|-------------------------------------------------------------------------|--------------------------------------------------------------------------------|--------------------------------------------------------|
| Viewer A | Positive | 0 | 0 | 0 | 13 | 26 |
| | Negative | 10 | 10 | 20 | 1 | 0 |
| Viewer B | Positive | 0 | 0 | 0 | 13 | 26 |
| | Negative | 10 | 10 | 20 | 1 | 0 |
| Viewer C | Positive | 0 | 0 | 0 | 13 | 26 |
| | Negative | 10 | 10 | 20 | 1 | 0 |
{18}------------------------------------------------
| Discordant Results | | | | | |
|--------------------|---------------|--------|------------------------------|--|--|
| Viewer | | | GC/MS (ng/mL) DipCard Format | | |
| | Sample Number | Result | Viewer Results | | |
| Viewer A | 94910807 | 334 | Negative | | |
| Viewer B | 94910628 | 352 | Negative | | |
| Viewer C | 94910445 | 341 | Negative | | |
| Cup<br>format | | Negative | Low<br>Negative<br>by GC/MS<br>(less than<br>-50%) | Near Cutoff<br>Negative by<br>GC/MS<br>(Between<br>-50% and<br>cut-off) | Near<br>Cutoff<br>Positive by<br>GC/MS<br>(Between<br>the cut-off<br>and +50%) | High<br>Positive by<br>GC/MS<br>(greater<br>than +50%) |
|---------------|----------|----------|----------------------------------------------------|-------------------------------------------------------------------------|--------------------------------------------------------------------------------|--------------------------------------------------------|
| Viewer A | Positive | 0 | 0 | 0 | 13 | 26 |
| | Negative | 10 | 10 | 20 | 1 | 0 |
| Viewer B | Positive | 0 | 0 | 0 | 13 | 26 |
| | Negative | 10 | 10 | 20 | 1 | 0 |
| Viewer C | Positive | 0 | 0 | 1 | 13 | 26 |
| | Negative | 10 | 10 | 19 | 1 | 0 |
#### Discordant Results
| Viewer | Sample Number | GC/MS (ng/mL)<br>Result | Cup Format<br>Viewer Results |
|----------|---------------|-------------------------|------------------------------|
| Viewer A | 94910807 | 334 | Negative |
| Viewer B | 94910095 | 339 | Negative |
| Viewer C | 94910628 | 352 | Negative |
| Viewer C | 94910808 | 270 | Positive |
#### i. Lay-user study
A lay user study was performed at three intended user sites with 280 lay persons testing the Butalbital devices. A total of 141 females and 139 males tested the Butalbital samples. They had diverse educational and professional backgrounds and ranged in age from 21 to > 50 years. Urine samples were prepared at the following concentrations; negative, +/-75%, +/-50%, +/-25% of the cutoff by spiking drugs into drug free-pooled urine specimens. The concentrations of the samples were confirmed by GC/MS. Each sample was aliquoted into individual containers and blind-labeled. Each participant was provided with the package insert, 1 blind labeled sample and a device. The results are summarized below.
## Comparison between GC/MS and Lay Person Results (Butalbital DipCard)
| % of Cutoff | Number | Butalbital Concentration | Lay person results | The percentage |
|-------------|--------|--------------------------|--------------------|----------------|
|-------------|--------|--------------------------|--------------------|----------------|
{19}------------------------------------------------
| | of<br>samples | by GC/MS<br>(ng/mL) | No. of<br>Positive | No. of<br>Negative | of correct<br>results<br>(%) |
|--------------|---------------|---------------------|--------------------|--------------------|------------------------------|
| -100% Cutoff | 20 | 0 | 0 | 20 | 100% |
| -75% Cutoff | 20 | 77 | 0 | 20 | 100% |
| -50% Cutoff | 20 | 156 | 0 | 20 | 100% |
| -25% Cutoff | 20 | 234 | 1 | 19 | 95% |
| +25% Cutoff | 20 | 390 | 19 | 1 | 95% |
| +50% Cutoff | 20 | 468 | 20 | 0 | 100% |
| +75% Cutoff | 20 | 547 | 20 | 0 | 100% |
Comparison between GC/MS and Lay Person Results (Butalbital Cup)
| % of Cutoff | Number<br>of<br>samples | Butalbital Concentration<br>by GC/MS<br>(ng/mL) | Lay person results | | The percentage<br>of correct<br>results<br>(%) |
|--------------|-------------------------|-------------------------------------------------|--------------------|--------------------|------------------------------------------------|
| | | | No. of<br>Positive | No. of<br>Negative | |
| -100% Cutoff | 20 | 0 | 0 | 20 | 100% |
| -75% Cutoff | 20 | 77 | 0 | 20 | 100% |
| -50% Cutoff | 20 | 156 | 0 | 20 | 100% |
| -25% Cutoff | 20 | 234 | 1 | 19 | 95% |
| +25% Cutoff | 20 | 390 | 20 | 0 | 100% |
| +50% Cutoff | 20 | 468 | 20 | 0 | 100% |
| +75% Cutoff | 20 | 547 | 20 | 0 | 100% |
Lay-users were also given surveys on the ease of understanding the package insert instructions. All lay users indicated that the device instructions can be easily followed. A Flesch-Kincaid reading analysis was performed on each package insert and the scores revealed a reading Grade Level of 7.
- j. Clinical Studies
Not applicable.
## First Sign® Drug of Abuse Morphine Test
Analytical Performance
- a. Precision
Precision studies were carried out for samples with concentrations of -100% cut-off. -50% cut-off, -25% cut-off, at the cut-off, +25% cut-off, +75% cut-off, +75% cut-off and +100% cut-off. These samples were prepared by spiking drug in negative urine samples. Each drug concentration was confirmed by GC/MS. All sample aliquots were blind-labeled and randomized by the person who prepared samples and did not take part in the sample testing. For each concentration, tests were performed two runs per day for 25 days by three different operators for
{20}------------------------------------------------
each format of devices. Different set of operators tested each format. The results obtained are summarized in the following tables:
| Drug | Result<br>-100%<br>Cut-off | -75%<br>Cut-off | -50%<br>Cut-off | -25%<br>Cut-off | Cut-off | +25%<br>Cut-off | +50%<br>Cut-off | +75%<br>Cut-off | +100%<br>Cut-off |
|---------------------|----------------------------|-----------------|-----------------|-----------------|---------|-----------------|-----------------|-----------------|------------------|
| Lot 1 | 50-/0+ | 50-/0+ | 50-/0+ | 50-/0+ | 2-/48+ | 50+/0- | 50+/0- | 50+/0- | 50+/0- |
| Lot 2 | 50-/0+ | 50-/0+ | 50-/0+ | 50-/0+ | 1-/49+ | 50+/0- | 50+/0- | 50+/0- | 50+/0- |
| Lot 3 | 50-/0+ | 50-/0+ | 50-/0+ | 50-/0+ | 2-/48+ | 50+/0- | 50+/0- | 50+/0- | 50+/0- |
| Morphine Cup Format | | | | | | | | | |
| | Result<br>100% | 75% | 50% | 25% | | +25% | +50% | +75% | +100% |
## Morphine Dip Card Format
| Result | -100%<br>Cut-off | -75%<br>Cut-off | -50%<br>Cut-off | -25%<br>Cut-off | Cut-off | +25%<br>Cut-off | +50%<br>Cut-off | +75%<br>Cut-off | +100%<br>Cut-off |
|--------|------------------|-----------------|-----------------|-----------------|---------|-----------------|-----------------|-----------------|------------------|
| Drug | | | | | | | | | |
| Lot 4 | 50-/0+ | 50-/0+ | 50-/0+ | 50-/0+ | 2-/48+ | 50+/0- | 50+/0- | 50+/0- | 50+/0- |
| Lot 5 | 50-/0+ | 50-/0+ | 50-/0+ | 50-/0+ | 3-/47+ | 50+/0- | 50+/0- | 50+/0- | 50+/0- |
| Lot 6 | 50-/0+ | 50-/0+ | 50-/0+ | 50-/0+ | 2-/48+ | 50+/0- | 50+/0- | 50+/0- | 50+/0- |
## b. Linearity
Not applicable.
# c. Stability
The devices are stable at 39-86 (4-30°C) for 24 months based on the accelerated stability study at 50°C. Control materials are not provided with the device. The labeling provides information on how to obtain control materials.
# d. Cut-off
A total of 150 samples equally distributed at concentrations of -50% cut-off; -25% cut-off; cut-off; +25% cut-off; +50% cut-off were tested using three different lots of each device by three different operators. Results were all positive at and above +25% cut-off and all negative at and below -25% cut-off for Morphine. The following cut-off value for the test devices have been verified.
| Test | Calibrator | Cut-off (ng/mL) |
|--------------------------------------------|------------|-----------------|
| First Sign® Drug of Abuse Morphine<br>Test | Morphine | 300 |
# e. Interference
Potential interfering substances found in human urine of physiological conditions were added to drug-free urine and to urine containing target drugs at 25% below and 25% above cut-off levels. These urine samples were tested using three lots of each device for all formats.
{21}------------------------------------------------
Compounds that showed no interference at a concentration of 100μg/mL are summarized in the following tables. There were no differences observed for different formats.
| Acebutolol | Ecgonine methylester | p-Hydroxymethamphetamine |
|----------------------|----------------------------------|------------------------------------------|
| Acetopromazine | (-) Y Ephedrine | Papaverine |
| 4-Acetamidophenol | Erythromycin | Penicillin-G |
| Acetophenetidin | β-Estradiol | Pentazocine |
| N-Acetylprocainamide | Estrone-3-sulfate | Pentobarbital |
| Acetylsalicylic acid | Ethyl-p-aminobenzoate | Perphenazine |
| Aminopyrine | Fenoprofen | Phencyclidine |
| Amitryptyline | Furosemide | Phenelzine |
| Amobarbital | Gentisic acid | Phenobarbital |
| Amoxicillin | Hemoglobin | Phentermine |
| Ampicillin | Hydralazine | L-Phenylephrine |
| Ascorbic acid | Hydrochlorothiazide | β-Phenylethlamine |
| Amphetamine | Hydrocortisone | β-Phenyllethylamine |
| L-Amphetamine | O-Hydroxyhippuric acid | Phenylpropanolamine |
| Apomorphine | 3-Hydroxytyramine | Prednisolone |
| Aspartame | Ibuprofen | Prednisone |
| Atropine | Imipramine | Promazine |
| Benzilic acid | Iprazid | Promethazine |
| Benzoic acid | (-) -Isoproterenol | D,L-Propanolol |
| Benzoylecgonine | Isoxsuprine | D-Propoxyphene |
| Benzphetamine | Ketamine | D-Pseudoephedrine |
| Bilirubin | Ketoprofen | Quinidine |
| Brompheniramine | Labetalol | Quinine |
| Buprenorphine | Loperamide | Ranitidine |
| Butalbital | Maprotiline | Salicylic acid |
| Caffeine | Meprobamate | Secobarbital |
| Chloralhydrate | Methadone | Serotonin<br>(5-Hydroxytyramine) |
| Chloramphenicol | Methamphetamine | Sulfamethazine |
| Chlordiazepoxide | Methoxyphenamine | Sulindac |
| Chlorothiazide | (+)3,4-Methylenedioxyamphetamine | Temazepam |
| (±) Chlorpheniramine | Methylenedioxymethamphetamine | Tetracycline |
| Chlorpromazine | Methylphenidate | Tetrahydrocortisone3(5-D<br>glucuronide) |
| Chlorquine | Nalorphine | Tetrahydrozoline |
{22}------------------------------------------------
| Cholesterol | Naloxone | Thiamine |
|--------------------------------------------------------|----------------------|-----------------|
| Clomipramine | Nalidixic acid | Thioridazine |
| Clonidine | Naltrexone | D, L-Thyroxine |
| Cocaine hydrochloride | Naproxen | Tolbutamine |
| Cortisone…
